摘要目的 观察原花青素(OPC)对脊髓损伤(SCI)小鼠的神经保护作用.方法 采用改良Allen's击打法制备SCI小鼠模型,将小鼠随机分为Vehicle(Veh)组和原青花素组.术后3d,二氢乙锭(HEt)染色检测活性氧(ROS)的积累,酶联免疫吸附试验(ELISA)试剂盒检测肿瘤坏死因子-α(TNF-α)和白细胞介素-1β(IL-1β)的表达,检测裂解的半胱氨酰天冬氨酸特异性蛋白酶-3(Cleaved Caspase-3)蛋白检测损伤周围坏死细胞数量;术后28 d,Basso Mouse Scale(BMS)评分检测小鼠后肢运动功能,悬尾实验评价小鼠情绪状况,髓磷脂碱性蛋白(MBP)检测白质损伤情况.结果 (1)BMS评分结果显示原青花素治疗组第7、14、21、28天后肢运动功能评分[(2.38±0.26)、(3.25±0.31)、(3.75±0.45)和(4.25±0.37)分]与Veh组比较,差异有统计学意义[(1.00±0.27)、(1.75±0.16)、(2.00±0.19)、(2.63±0.26)分,P=0.010].(2)悬尾实验表明原花青素治疗组和Veh组比较差异有统计学意义[(248.33±2.65)s和(281.33±3.853)s,t=7.053,P=0.000].(3)对比Veh组,原青花素组抑炎因子IL-1β表达量[(719.5±55.9) ng/ml]明显低于Veh组[(1 100.5±61.5) ng/ml,=4.583,P=0.001],TNF-α[(829.2±50.9) ng/ml]明显低于Veh组[(1 113.0±58.8)ng/ml,=3.648,P=0.005].(4)原青花素组ROS的荧光强度(1 154.8±78.5)明显低于Veh组(1 689.0±86.9),差异有统计学意义(t=4.562,P=0.000).(5)对比Veh组[(18.7±1.5)％],原青花素组的正常髓鞘明显增多[(24.5±1.8)％],差异有统计学意义(t=2.474,P=0.033).(6)对比Veh组,原花青素组Cleaved Caspase-3的表达量下调(0.342±0.028和0.156±0.036,t=4.053,P=0.002).结论 原青花素对脊髓损伤小鼠的神经保护作用.

abstractsObjective To investigate the neuroprotective effect of proantho cyanidins (OPC) on spinal cord injury (SCI) mice.Methods The SCI mouse model was prepared by modified Allen' s hitting method.At 3 days after SCI,hydroethidine (HEt) staining was used to detect reactive oxygen species (ROS) production.Enzyme linked immunosorbent assay (ELISA) kit was used to detect the expression of tumor necrosis factor-α (TNF-α) and interleukin (IL)-1 β.The expression of Cleaved cysteinyl aspartate-specific protease (Caspase)-3 was detected by Western blotting.At 28 days after SCI,Basso Mouse Scale (BMS) score was used to detect the hindlimb motor function.Tail suspension test was used to evaluate the depressive status of mice.The expression of myelin basic protein (MBP) was detected by immunofluorescence on day 28 post injury.Results (1) BMS scores showed that OPC treatment group 7,14,21 and 28 d scores of hindlimb locomotor function (2.38 ± 0.26,3.25 ± 0.31,3.75 ± 0.45 and 4.25 ±0.37) compared with the Veh Group (1.00 ±0.27,1.75 ±0.16,2.00 ±0.19 and 2.63 ±0.26,P =0.010).(2) Tail suspension test showed that the difference between the OPC treatment group and the Veh group was statistically significant [(248.33 ± 2.65) s and (281.33 ± 3.853) s,t =7.053,P =0.000].(3) Compared with Veh group,OPC group of proinflammatory factor IL-1β was obviously lower [(719.5±55.9) ng/ml vs.(1 100.5±61.5) ng/ml,t=4.583,P=0.001].TNF-α [(829.2 ± 50.9) ng/ml] was significantly lower than that of Veh group [(1 113.0 ±58.8) ng/ml,t =3.648,P =0.005].(4) The fluorescence intensity of ROS in OPC group (1 377.00±27.06) was significantly lower than Veh group (1 689.0 ± 8.0,t =4.562,P =O.000).(5) Compared with Veh group [(18.7 ± 1.5) ％],the percent of MBP positive area in OPC group was significantly increased [(24.5 ± 1.8) ％,t =2.474,P =0.033].(6) Compared with Veh group,the expression of Cleaved Caspase-3 in OPC group was decrease (0.342 ± 0.028 vs.0.156 ± 0.036,t =4.053,P =0.002).Conclusion OPC protects neurons in spinal cord.